http://confocal-microscopy-list.275.s1.nabble.com/Cy7-TSA-dye-tp7587083p7587091.html
Thanks for the info. I am aware of the paper that you mentioned. They use a
Yes, spectral imaging is the way to go but it has its own challenges. I
> *****
> To join, leave or search the confocal microscopy listserv, go to:
>
http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy> Post images on
http://www.imgur.com and include the link in your posting.
> *****
>
> Hi Sripad,
>
> Molecular Probes can conjugate whatever Alexa Fluor you want onto
> tyramide, and yes, there is an AF Cy7 equivalent.Tyramide is off patent,
> other 'chemistry' companies could conjugate their/your favorite
> fluorophores.
>
> //
>
> At higher plex, you are going to have spectral overlap: learn to love
> spectral unmixing. For example:
>
> PLoS One. 2016 Jul 8;11(7):e0158495. doi: 10.1371/journal.pone.0158495.
> eCollection 2016.
>
> Multiplexed Spectral Imaging of 120 Different Fluorescent Labels.
>
> Valm AM(1)(2), Oldenbourg R(1)(2), Borisy GG(3).
>
> Author information:
> (1)Marine Biological Laboratory, Woods Hole, Massachusetts, United States
> of
> America.
> (2)Brown University, Providence, Rhode Island, United States of America.
> (3)Forsyth Institute, Cambridge, Massachusetts, United States of America.
>
> The number of fluorescent labels that can unambiguously be distinguished
> in a
> single image when acquired through band pass filters is severely limited
> by the
> spectral overlap of available fluorophores. The recent development of
> spectral
> microscopy and the application of linear unmixing algorithms to spectrally
> recorded image data have allowed simultaneous imaging of fluorophores with
> highly
> overlapping spectra. However, the number of distinguishable fluorophores
> is still
> limited by the unavoidable decrease in signal to noise ratio when
> fluorescence
> signals are fractionated over multiple wavelength bins. Here we present a
> spectral image analysis algorithm to greatly expand the number of
> distinguishable
> objects labeled with binary combinations of fluorophores. Our algorithm
> utilizes
> a priori knowledge about labeled specimens and imposes a binary label
> constraint
> on the unmixing solution. We have applied our labeling and analysis
> strategy to
> identify microbes labeled by fluorescence in situ hybridization and here
> demonstrate the ability to distinguish 120 differently labeled microbes in
> a
> single image.
>
> DOI: 10.1371/journal.pone.0158495
> PMCID: PMC4938436
> PMID: 27391327
>
>
>
> George
>
>
> On 7/28/2017 2:22 PM, S Ram wrote:
>
>> *****
>> To join, leave or search the confocal microscopy listserv, go to:
>>
http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy>> Post images on
http://www.imgur.com and include the link in your posting.
>> *****
>>
>>
>> Hello,
>> Just wondering if anyone has ever come across (or used) a Cy7 or similar
>> TSA dye probe? I checked a few of the vendors that I could think of
>> (Molecular Probes/Invitrogen/Fisher, Perkin Elmer) but none of them had
>> any
>> probes in that range.
>>
>> The goal is to do automated staining of FFPE tissues using Cy5 and another
>> NIR probe. The dyes need to be TSA conjugates. I do not want to use Cy5.5
>> due to spectral overlap. Also, dyes in the visible region are no beuno due
>> to too much autofluorescence.
>>
>> Any help/suggestion is greatly appreciated.
>>
>> Thanks.
>>
>> Sripad
>>
>
> --
>
>
> George McNamara, PhD
> Baltimore, MD 21231
>
[hidden email]
>
https://www.linkedin.com/in/georgemcnamara>
https://works.bepress.com/gmcnamara/75 (may need to use Microsoft Edge
> or Firefox, rather than Google Chrome)
>
http://www.ncbi.nlm.nih.gov/myncbi/browse/collection/44962650>
http://confocal.jhu.edu (as of May 22, 2017)
>